Human papillomaviruses have been implicated as etiologic agents in cervical cancer. The transforming protein, E6 and E7 of oncogenic HPVs are consistently expressed in tumor cells and therefore represent examples of tumor specific antigens. Vaccines targeted to these proteins may provide a means to prevent and treat HPV associated malignancies. In order to design and characterize the efficacy of various vaccine strategies and other immunotherapeutic strategies, it will be important to characterize antigen specific T cell responses to the HPV E6 and E7 proteins. Over the past few years, our group has developed novel genetic approaches to cancer immunotherapy which result in the induction or enhancement of both CTL and T helper responses to tumor specific antigens. As HPV associated malignancies represent one of the best models for human cancer in which there are identified tumor specific antigen, we propose to study these immune responses and characterize specific E6 and E7 epitopes that are expressed in patients bearing the most common MHC class I allele - HLA A2. Specifically, we propose to: 1) develop and utilize assays to measure CTL and Th responses to HPV 16 E6 and E7 proteins. A variety of strategies will be considered to prepare stimulator and target cells for CTL and Th assays. 2) identify epitopes on HPV 16 E6 and E7 proteins which bind to commonly expressed human HLA class I alleles and generate CTL responses. Naturally processed HPV 16 E6 and E7 peptides will be characterized by immunoprecipitation of MHC class I-peptide complexes from HPV-associated cervical carcinoma. Acid dissociated peptides will be analyzed by HPLC and compared with the elution profiles of synthetic peptides. 3) measure and characterize CTL and Th responses to HPV 16 E6 and E7 proteins/peptides in peripheral blood of patients treated with cytokine gene transduced tumor cell vaccines or with recombinant vaccinia-E6/E7 vaccines.